Gene modulation for protein ZO-3 and its effects on urothelial barrier formation, function and recovery. (360G-Wellcome-211684_Z_18_Z)

The question I propose to address is what role ZO-3 plays in urothelial barrier function and repair. I will tackle this by assessing how ZO-3 knockdown influences urothelial barrier tightness and the speed of urothelial repair. I will conduct various experiments to prove this including; CRISPR-Cas (or shRNA as contingency) gene deletion, immunoblotting (claudin3 analysis) and TER for measuring the strength of the urothelial barrier. I predict that ZO3 inhibition (knockdown) will result in a weaker barrier but an increase in recovery time. I think the loss of ZO-3 may increase recovery time as it may aid in recruiting key repair machinery involved in reforming the tight junction, therefore its loss would slow the time it takes to repair. This, in turn, may shine a light on how bladder cancer may be able to metastasize due to weak urothelial barriers, caused by mutations in ZO3 gene and become a new therapeutic target for malignant bladder cancer.